Orthogonal and multiplexable genetic perturbations with an engineered prime editor and a diverse RNA array

• 全文链接

通过工程化的原始编辑器和多样化的RNA阵列进行正交和可多重化的基因干扰

关键词：

来源：

Nature Communications

全文链接：

//agri.nais.net.cn/topic/downloadFile/ac242082-5980-4855-aafc-f6d9429d386a

来源地址：

https://www.nature.com/articles/s41467-024-55134-9

资源所属：

马铃薯基因组设计育种

类型：

学术文献

语种：

中文

原文发布日期：

2025-01-09

摘要：

Programmable and modular systems capable of orthogonal genomic and transcriptomic perturbations are crucial for biological research and treating human genetic diseases. Here, we present the minimal versatile genetic perturbation technology (mvGPT), a flexible toolkit designed for simultaneous and orthogonal gene editing, activation, and repression in human cells. The mvGPT combines an engineered compact prime editor (PE), a fusion activator MS2–p65–HSF1 (MPH), and a drive-and-process multiplex array that produces RNAs tailored to different types of genetic perturbation. mvGPT can precisely edit human genome via PE coupled with a prime editing guide RNA and a nicking guide RNA, activate endogenous gene expression using PE with a truncated single guide RNA containing MPH-recruiting MS2 aptamers, and silence endogenous gene expression via RNA interference with a short-hairpin RNA. We showcase the versatility of mvGPT by simultaneously correcting a c.3207C>A mutation in the ATP7B gene linked to Wilson’s disease, upregulating the PDX1 gene expression to potentially treat Type I diabetes, and suppressing the TTR gene to manage transthyretin amyloidosis. In addition to plasmid delivery, we successfully utilize various methods to deliver the mvGPT payload, demonstrating its potential for future in vivo applications.